福建省彩叶草叶斑病病原菌的分离鉴定与室内药剂筛选

王荣波; 陈姝樽; 李本金; 刘裴清; 邓丽霞; 翁启勇

doi:10.19303/j.issn.1008-0384.2021.12.011

福建省彩叶草叶斑病病原菌的分离鉴定与室内药剂筛选

doi: 10.19303/j.issn.1008-0384.2021.12.011

福建省农业科学院植物保护研究所/福建省作物有害生物监测与治理重点实验室，福建　福州　 350013

基金项目: 福建省科技计划公益类专项（2019R1024-4）；福建省自然科学基金项目（2021J01478）；福建省农业科学院科技创新团队建设项目（CXTD2021002-1）；福建省农业高质量发展超越“5511”协同创新工程项目(XTCXGC2021017、 XTCXGC2021011); 福建省农业科学院对外合作项目（DWHZ2021-13）

详细信息

作者简介:
王荣波（1988−），男，博士，助理研究员，研究方向：植物病理学（E-mail：wrb16128@163.com）

通讯作者:
翁启勇（1962−），男，硕士，研究员，研究方向：作物有害生物综合治理技术研究和应用（E-mail： wengqy@faas.cn）

中图分类号: S 436
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- 被引次数: 0
出版历程
- 收稿日期: 2021-11-15
- 修回日期: 2021-11-27
- 网络出版日期: 2021-12-30
- 刊出日期: 2021-12-28

Pathogen Identification and Fungicide Selection for Leaf Spot Disease on Plectranthus scutellarioides in Fujian

Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests/Institute of Plant Protection, Fuzhou, Fujian　350013, China

摘要

摘要: 目的明确福建省彩叶草（Plectranthus scutellarioides）叶部病害的病原种类，筛选可用于病害防控的化学药剂。方法采用常规组织分离法获得病原菌株并进行柯赫氏法则验证，结合形态学与分子生物学（ITS与LSU序列）对分离菌株进行鉴定，采用生长速率法测定6种杀菌剂对该病原菌的室内毒力。结果用分离菌株ZZCYC1706分生孢子接种彩叶草，侵染叶片症状与田间症状相似；形态特征及ITS和LSU多基因序列分析表明，该病原菌为链格孢（Alternaria alternata）。供试6种药剂室内毒力测定结果显示，96%啶菌恶唑抑菌效果最好，其EC₅₀为0.621 μg·mL⁻¹，其次为98%异菌脲和95%嘧霉胺，EC₅₀分别为1.155 、5.258 μg·mL⁻¹；而96.4%克菌丹效果较差，EC₅₀值最高，为89.010 μg·mL⁻¹。结论福建漳州彩叶草叶斑病病原菌鉴定为链格孢（A. alternata），该发现为国内首次报道。啶菌恶唑、异菌脲和嘧霉胺对该病原菌的毒力较高，并分属不同药剂类型，可用于链格孢引起叶斑病的防治。
- 彩叶草 /
- 叶斑病 /
- 链格孢 /
- 毒力测定 /
- EC₅₀
Abstract: Objective Identification of the leaf spot pathogen on Plectranthus scutellarioides in Fujian and screening fungicides for effective disease prevention and control were conducted. Method A causative pathogen was isolated from the diseased tissues, and the species verified by Koch’s rule, morphological observations under a microscope, and molecular ITS and LSU sequence analyses. An in vitro toxicity test on 6 commercially available fungicides was performed on the pathogen to determine their efficacy. Result The coleus leaves inoculated with the conidia of the isolated ZZCYC1706 produced same symptoms as those observed on the diseased plants in the field. The ITS and LSU sequences of ZZCYC1706 suggested it be Alternaria alternata. The toxicity test showed, among the 6 fungicides, 96% pyrisoxazole to be most effective in inhibiting the pathogenic growth with the lowest EC₅₀ of 0.621 μg·mL⁻¹. It was followed by 98% iprodione with an EC₅₀ at 1.155 μg·mL⁻¹ , 95% pyrimethanil at 5.258 μg·mL⁻¹, and 96.4% captan, which was the least effective, at 89.010 μg·mL⁻¹. Conclusion The pathogen of the leaf spot disease on P. scutellarioides in Fujian province was identified as A. alternata, which was the first reported in China. The fungicides, pyrisoxazole, iprodione, and pyrimethanil, were shown in the laboratory effective in inhibiting the growth of A. alternata. And, being of different types, these fungicides could potentially be applicable for the coleus leaf spot disease control with a reduced concern of the development of drug resistance by the pathogen.
- Plectranthus scutellarioides /
- leaf spot /
- Alternaria alternata /
- toxicity test /
- EC₅₀

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图 1 彩叶草叶斑病自然发病症状

注：A，田间植株发病状况；B，植株叶片发病状况；C，正面病斑；D，背面病斑

Figure 1. Leaf spots on coleus occurred in nature

Note: A: Infected leaves on field plants; B: Typical symptoms on diseased plant; C: Disease spots on face of leaf; D: Disease spots on back of leaf.

下载: 全尺寸图片幻灯片

图 2 菌落形态和致病性鉴定

注：A，生长3 d的菌落形态；B，生长10 d的正面菌落形态；C，生长10 d的反面菌落形态；D，接种对照叶片（3 d）；E，分生孢子接种叶片（3 d）

Figure 2. Colony morphology and pathogenic assay

Note: A: Colony on PDA after 3 d incubation; B: Colony top-view after 10 d incubation; C: Colony bottom-view after 10 d incubation; D: Control leaf after 3 d incubation; E: Artificially inoculated leaf after 3 d incubation.

下载: 全尺寸图片幻灯片

图 3 病原菌分生孢子梗及分生孢子形态特征

注：A，早期分生孢子与分生孢子梗；B，分生孢子与有隔膜的菌丝；C，不同类型的分生孢子；标尺为50 μm

Figure 3. Morphological characteristics of conidiophore and conidia

Note: A: Early conidia with conidiophore; B: Conidia with septate mycelium hyphae; C: Different types of conidia; scale bar=50 μm.

下载: 全尺寸图片幻灯片

图 4 基于ITS和LSU序列构建链格孢属系统发育树

注：分支上的数字标示Bootstrap 检验的支持率，大丽轮枝菌（V. dahliae）作为外群，ZZCYC1706为分离菌株

Figure 4. Phylogenetic tree of Alternaria spp. based on ITS and LSU sequences

Note: Data shown at each node are confidence values from 1 000 replicate bootstrap samplings; Verticillium dahliae was out group, while ZZCYC1706, isolate.

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表 1 6种药剂对彩叶草叶斑病病原菌的毒力测定

Table 1. Toxicities of 6 fungicides on A. alternata

药剂 Fungicide	回归方程 Regression equation	相关系数r Correlation coefficient, r	EC₅₀值 EC ₅₀Value/ (µg·mL⁻¹)
95%嘧霉胺 95%Pyrimethanil	y=0.994x+3.647	0.985	5.258
98%异菌脲 98% Iprodione	y=0.827x+4.962	0.984	1.155
96.4%克菌丹 96.4% Captan	y=0.427x+3.085	0.917	89.010
96%啶菌恶唑 96% Pyrisoxazole	y=0.386x+5.184	0.999	0.621
99%腐霉利 99% Procymidone	y=0.535x+2.929	0.995	48.120
96%啶酰菌胺 96% Boscalid	y=0.566x+2.884	0.998	42.210

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